Profiling Genome-Wide Chromatin Methylation with Engineered Posttranslation Apparatus within Living Cells

详细信息    查看全文

• 作者：Rui Wang ; Kabirul Islam ; Ying Liu ; Weihong Zheng ; Haiping Tang ; Nathalie Lailler ; Gil Blum ; Haiteng Deng ; Minkui Luo
• 刊名：The Journal of the American Chemical Society
• 出版年：2013
• 出版时间：January 23, 2013
• 年：2013
• 卷：135
• 期：3
• 页码：1048-1056
• 全文大小：392K
• 年卷期：v.135,no.3(January 23, 2013)
• ISSN：1520-5126

文摘

Protein methyltransferases (PMTs) have emerged as important epigenetic regulators in myriad biological processes in both normal physiology and disease conditions. However, elucidating PMT-regulated epigenetic processes has been hampered by ambiguous knowledge about in vivo activities of individual PMTs particularly because of their overlapping but nonredundant functions. To address limitations of conventional approaches in mapping chromatin modification of specific PMTs, we have engineered the chromatin-modifying apparatus and formulated a novel technology, termed clickable chromatin enrichment with parallel DNA sequencing (CliEn-seq), to probe genome-wide chromatin modification within living cells. The three-step approach of CliEn-seq involves in vivo synthesis of S-adenosyl-l-methionine (SAM) analogues from cell-permeable methionine analogues by engineered SAM synthetase (methionine adenosyltransferase or MAT), in situ chromatin modification by engineered PMTs, subsequent enrichment and sequencing of the uniquely modified chromatins. Given critical roles of the chromatin-modifying enzymes in epigenetics and structural similarity among many PMTs, we envision that the CliEn-seq technology is generally applicable in deciphering chromatin methylation events of individual PMTs in diverse biological settings.